Non-specific phospholipase C4 hydrolyzes phosphosphingolipids and phosphoglycerolipids and promotes rapeseed growth and yield

Phosphorus is a major nutrient vital for plant growth and development,with a substantial amount of cellular phosphorus being used for the biosynthesis of membrane phospholipids.Here,we report that NON-SPECIFIC PHOSPHOLIPASE C4(NPC4)in rapeseed(Brassica napus)releases phosphate from phospholipids to promote growth and seed yield,as plants with altered NPC4 levels showed significant changes in seed production under different phosphate conditions.Clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated nuclease 9(Cas9)-mediated knockout of Bna NPC4 led to elevated accumulation of phospholipids and decreased growth,whereas overexpression(OE)of Bna NPC4resulted in lower phospholipid contents and increased plant growth and seed production.We demonstrate that Bna NPC4 hydrolyzes phosphosphingolipids and phosphoglycerolipids in vitro,and plants with altered Bna NPC4 function displayed changes in their sphingolipid and glycerolipid contents in roots,with a greater change in glycerolipids than sphingolipids in leaves,particularly under phosphate deficiency conditions.In addition,Bna NPC4-OE plants led to the upregulation of genes involved in lipid metabolism,phosphate release,and phosphate transport and an increase in free inorganic phosphate in leaves.These results indicate that Bna NPC4 hydrolyzes phosphosphingolipids and phosphoglycerolipids in rapeseed to enhance phosphate release from membrane phospholipids and promote growth and seed production.